scholarly journals Validation of a method of broth microdilution for the determination of antibacterial activity of essential oils

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Vanegas David ◽  
Abril-Novillo Andrea ◽  
Khachatryan Aleksandr ◽  
Jerves-Andrade Lourdes ◽  
Peñaherrera Eugenia ◽  
...  
Keyword(s):  
Essential Oils ◽  
Sensitivity And Specificity ◽  
Inhibitory Concentration ◽  
Tween 80 ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Broth Microdilution ◽  
Continuous Contact ◽  
Microdilution Method ◽  
Broth Microdilution Method

Abstract Objective The aim of the present study was to adapt and optimize a broth microdilution method and compare it to the agar dilution method for the evaluation of activity of essential oils from medicinal plants against Gram-negative bacteria. Based on bibliographic research, active and not active oils were selected. The sensitivity and specificity were established as parameters for validation. The comparison between both methods was made using contingency analysis tables, based on the observed frequencies. For both methods, the minimum inhibitory concentration was determined against Escherichiacoli strains, in an essential oil concentration range between 0.03 and 0.48% (v/v). Results A stable emulsion formation was achieved with the addition of Tween 80 and constant agitation, guaranteeing the continuous contact of oil with bacteria (critical step in the microdilution method). The statistical analysis of results obtained with both methods presented a good sensitivity and specificity (100% in both cases), which let us correctly discriminate between active and non-active oils. The values obtained for the minimal inhibitory concentration were independent of the technique used. Finally, the obtained results show that the validated microtechnique allows important diminishment of time and resources for investigations dealing with essential oils or lipophilic extracts evaluation.

scholarly journals Comparison of Broth Microdilution Method UsingHaemophilus Test Medium and Agar Dilution Method for Susceptibility Testing of Eikenella corrodens

1998 ◽  
Vol 36 (8) ◽  
pp. 2386-2388 ◽  
Author(s):  
Luis Alcalá ◽  
Fernando García-Garrote ◽  
Emilia Cercenado ◽  
Teresa Peláez ◽  
Gema Ramos ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Broth Microdilution ◽  
Test Medium ◽  
Eikenella Corrodens ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Agar Dilution

Susceptibility testing of Eikenella corrodens is usually performed by a Mueller-Hinton sheep blood agar dilution (AD) method. However, this method is impractical for testing only a few strains. We compared AD with the broth microdilution method usingHaemophilus test medium (HTM) in order to determine the susceptibility of 36 clinical isolates of E. corrodens to eight antimicrobial agents. MICs obtained by the HTM method yielded 95.5 and 84% agreement (within 2 and 1 log2 dilutions, respectively) with those obtained by AD. The HTM method with incubation in CO2 for 48 h was highly reproducible and constitutes an easy alternative for antimicrobial susceptibility testing of E. corrodens.

scholarly journals Examination of the minimum inhibitory concentration of amoxicillin and marbofloxacin against Streptococcus suis using standardised methods

2020 ◽  
Vol 65 (No. 9) ◽  
pp. 377-386
Author(s):  
D Sperling ◽  
H Karembe ◽  
M Zouharova ◽  
K Nedbalcova
Keyword(s):  
Inhibitory Concentration ◽  
Streptococcus Suis ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Microdilution Method ◽  
Linear Relationships ◽  
Broth Microdilution Method ◽  
Significant Difference ◽  
Agar Dilution ◽  
Good Agreement

The results of the antimicrobial susceptibility testing of clinical isolates Streptococcus suis to amoxicillin and marbofloxacin obtained by the agar dilution method and broth microdilution method with the results obtained by the commercially available E-test were compared. Comparisons between the methods based on the determination of the minimal inhibitory concentration (MIC) of the antimicrobials were assessed based on the degree and frequency of the categorical agreement (Agar dilution method as a reference system) and the percentage of the categorical agreement and error rate. A statistical evaluation was determined using the Bland-Atman method. The presented MIC values, determined for the isolates in the E-test, were slightly different from the MIC values determined by the dilution tests, mainly due to the different defined testing concentrations. For the E-test as the test system and agar-dilution method as the reference system, no error of any class was detected (very major, major and minor error) and a complete categorical agreement was obtained between the evaluated methods for amoxicillin. For amoxicillin, the regression and correlation analysis show linear relationships between the E-test and the two dilution methods with significant coefficients of determination (0.62 and 0.75). The slopes of the equality and regression lines were not significantly different. However, the E-test tends to slightly overestimate the MIC values when compared to the microdilution. The reverse is true when compared with the agar dilution. There was good agreement between the E-test and the dilution methods with a low bias (0.001 3 and −0.005 0), all the experimental data were within the computed limits of agreement. For marbofloxacin, the same trends were observed with lower coefficients of determination (0.42 and 0.73) and a less favourable agreement. The E-test constantly underestimated the MIC values when compared to the two dilution methods. No significant difference between the microdilution and agar dilution was obtained.

Antimicrobial Essential Oil Combinations to Combat Foot Odour

Planta Medica ◽  
2018 ◽  
Vol 84 (09/10) ◽  
pp. 662-673 ◽  
Author(s):  
Ané Orchard ◽  
Alvaro Viljoen ◽  
Sandy van Vuuren
Keyword(s):  
Antimicrobial Activity ◽  
Essential Oil ◽  
Essential Oils ◽  
Antimicrobial Efficacy ◽  
Juniperus Virginiana ◽  
Broth Microdilution ◽  
Synergistic Interactions ◽  
Microdilution Method ◽  
Broth Microdilution Method

AbstractFoot odour (bromodosis) is an embarrassing and perplexing condition mostly caused by bacteria of the Brevibacterium species. Essential oils are a credible option as an affordable treatment of odour and contribute towards antimicrobial efficacy. Therefore, this study sets out to investigate the antimicrobial activity of essential oil combinations against odour-causing bacteria. The broth microdilution method was used to investigate the antimicrobial activity of 119 essential oil combinations, and the fractional inhibitory index was calculated to determine the interactive profile. Combinations that resulted in synergy in 1 : 1 ratios were further evaluated in different concentrations, and isobolograms were plotted to determine the influence of the ratio on overall activity. Numerous combinations could be identified as having synergistic interactions against the Brevibacterium spp. and no antagonism was observed. The combination of Juniperus virginiana (juniper) and Styrax benzoin (benzoin) demonstrated synergy against all three Brevibacterium spp. tested and J. virginiana was the essential oil responsible for the majority of the synergistic interactions. The results reported here confirm the promising potential of the majority of these oils and selected combinations in treating and controlling bromodosis.

scholarly journals In VitroAntistaphylococcal Effects ofEmbelia schimperiExtracts and Their Component Embelin with Oxacillin and Tetracycline

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Johana Rondevaldova ◽  
Olga Leuner ◽  
Alemtshay Teka ◽  
Ermias Lulekal ◽  
Jaroslav Havlik ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Medicinal Plants ◽  
Bacterial Infections ◽  
Inhibitory Concentration ◽  
Developed Countries ◽  
Positive Interactions ◽  
Fractional Inhibitory Concentration ◽  
Broth Microdilution ◽  
Microdilution Method ◽  
Broth Microdilution Method

Bacterial infections are in less-developed countries traditionally treated by remedies prepared from medicinal plants.Embelia schimperi(Vatke) is a plant used as a taenicide or disinfectant in Ethiopia, very often taken mixed with another plant species. In the present study, we examined two extracts prepared from seeds and twigs with leaves ofE. schimperiand its main present secondary metabolite embelin for their antibacterial combinatory effect with oxacillin and tetracycline against sensitive and resistantStaphylococcus aureusstrains. Minimum inhibitory concentrations were determined through the broth microdilution method, whereas the combinatory effect was evaluated through fractional inhibitory concentration sum (ΣFIC) indices. Results show many positive interactions and synergy occurring in embelin and oxacillin combinations against 4 out of 9 strains (ΣFIC 0.203–0.477) and for embelin and tetracycline combination against 3 out of 9 strains (ΣFIC 0.400–0.496). Moreover, the resistance to oxacillin has been overcome in 2 strains and to tetracycline in 3 strains. According to our knowledge, this is the first study showing antimicrobial combinatory effect ofE. schimperias well as of embelin. These findings can be used for the further research targeted on the development of new antistaphylococcal agents.

scholarly journals In Vitro Interaction of Caspofungin Acetate with Voriconazole against Clinical Isolates of Aspergillus spp

2002 ◽  
Vol 46 (9) ◽  
pp. 3039-3041 ◽  
Author(s):  
Sofia Perea ◽  
Gloria Gonzalez ◽  
Annette W. Fothergill ◽  
William R. Kirkpatrick ◽  
Michael G. Rinaldi ◽  
...  
Keyword(s):  
Inhibitory Concentration ◽  
Fractional Inhibitory Concentration ◽  
Broth Microdilution ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
In Vitro Interaction ◽  
Vitro Data ◽  
In Vitro Data

ABSTRACT The interaction between caspofungin acetate and voriconazole was studied in vitro by using 48 clinical Aspergillus spp. isolates obtained from patients with invasive aspergillosis. MICs were determined by the NCCLS broth microdilution method. Synergy, defined as a fractional inhibitory concentration (FIC) index of <1, was detected in 87.5% of the interactions; an additive effect, defined as an FIC index of 1.0, was observed in 4.2% of the interactions; and a subadditive effect, defined as an FIC index of 1.0 to 2.0, was found in 8.3% of the interactions. No antagonism was observed. Animal models are required to validate the in vivo significance of these in vitro data presented for the combination of caspofungin and voriconazole.

scholarly journals In VitroActivity of Fosfomycin against a Collection of Clinical Pseudomonas aeruginosa Isolates from 16 Spanish Hospitals: Establishing the Validity of Standard Broth Microdilution as Susceptibility Testing Method

2013 ◽  
Vol 57 (11) ◽  
pp. 5701-5703 ◽  
Author(s):  
María Díez-Aguilar ◽  
María-Isabel Morosini ◽  
Rosa del Campo ◽  
María García-Castillo ◽  
Javier Zamora ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Susceptibility Testing ◽  
Disk Diffusion ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Broth Microdilution ◽  
Content Type ◽  
Testing Method ◽  
Broth Microdilution Method ◽  
Agar Dilution

ABSTRACTThe broth microdilution method for fosfomycin andPseudomonas aeruginosawas assessed and compared with the approved agar dilution method in 206 genetically unrelatedP. aeruginosaclinical isolates. Essential agreement between the two methods was 84%, and categorical agreement was 89.3%. Additionally, Etest and disk diffusion assays were performed. Results validate broth microdilution as a reliable susceptibility testing method for fosfomycin againstP. aeruginosa. Conversely, unacceptable concordance was established between Etest and disk diffusion results with agar dilution results.

scholarly journals Antimicrobial resistance of Enterococcus faecium and Enterococcus faecalis, isolated from blood culture of patients with hematological malignancies during different study periods

2021 ◽  
Vol 16 (1) ◽  
pp. 54-63
Author(s):  
A. V. Fedorova ◽  
G. A. Klyasova ◽  
I. N. Frolova ◽  
S. A. Khrulnova ◽  
A. V. Vetokhina ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Blood Culture ◽  
Enterococcus Faecalis ◽  
Enterococcus Faecium ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
High Susceptibility ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
High Level

Objective: to determine antimicrobial resistance of Enterococcus faecium and Enterococcus faecalis isolated from blood culture of hematological patients during different study periods.Materials and methods. Antimicrobial susceptibility of Enterococcus spp., collected as part of the multicenter study was tested by the broth microdilution method (USA Clinical and Laboratory Standards Institute (CLSI), 2018), to daptomycin by Etest (bioMeriéux, France). High-level gentamicin resistance (HLGR) and high-level streptomycin resistance (HLSR) was performed by the agar dilution method (CLSI (Oxoid, UK), 2018).Results. The susceptibility of 366 E. faecium (157 in 2002-2009 and 209 in 2010-2017) and 86 E. faecalis (44 in 20022009 and 42 in 2010-2017) was studied. In the second study period (2010-2017) the rise of vancomycin-resistant E. faecium (VREF) increased from 8.3 % to 23.4 % (p = 0.0001), and two linezolid-resistant (LREF) were identified. All VREF and LREF remained susceptible to daptomycin and tigecycline. The rate of susceptible to tetracycline E. faecium remained the same (73.9 and 74.6 %), and an increase in susceptibility to chloramphenicol (74.5 and 82.3 %) was observed. Susceptibility of E. faecium to tetracycline was detected with almost the same rate and in a part of isolates, the increase of susceptibility to chloramphenicol was registered during the analyzed periods. The rise of E. faecium susceptible to HLGR and HLSR has increased significantly in 2010-2017 compared to 2002-2009. Erythromycin, levofloxacin, ampicillin and penicillin had the least activity against E. faecium (less than 5 %).All E. faecalis were susceptible to tigecycline, linezolid, and teicoplanin. Only one of E. faecalis had intermediate resistance to vancomycin. High susceptibility to ampicillin in E. faecalis remained unchanged (97.7 and 97.6 %, respectively). In the second period of the study the rise of susceptible E. faecalis decreased significantly to penicillin (from 97.7 % to 76.2 %), to levofloxacin (from 59.1 % to 31 %), to HLSR (from 52.3 % до 31 %), and to HLGR (from 47.7 % to 26.2 %), remained unchanged to chloramphenicol (52.3 % and 50 %) and was minimal to erythromycin and tetracycline.Conclusion. The study demonstrated higher rates of antibiotic resistance among E. faecium, which consisted of an increase in VREF and the appearance of linezolid-resistant strains. High susceptibility to ampicillin remained in E. faecalis, but there was an increase in resistance to penicillin and aminoglycosides.

Chemical composition and antimicrobial activity of Erodium species: E. ciconium L., E. cicutarium L., and E. absinthoides Willd. (Geraniaceae)

2010 ◽  
Vol 64 (3) ◽  
Author(s):  
Zorica Stojanović-Radić ◽  
Ljiljana Čomić ◽  
Niko Radulović ◽  
Milan Dekić ◽  
Vladimir Ranđelović ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Chemical Composition ◽  
Essential Oils ◽  
Palmitic Acid ◽  
Gram Negative Bacteria ◽  
Broth Microdilution ◽  
Bacterial Strains ◽  
Gram Negative ◽  
Microdilution Method ◽  
Broth Microdilution Method

AbstractThe present study gives results of chemical composition analyses and antimicrobial activity testing of three Erodium species: E. ciconium L., E. cicutarium L., and E. absinthoides Willd. Essential oils were obtained by hydro-distillation from air-dried entire plants and analyzed by GC and GC-MS. A total of 209 different compounds were identified: 162 for E. cicutarium, 107 for E. ciconium, and 79 for E. absinthoides. Antimicrobial activity (broth microdilution method) of the oils was screened against a panel of Gram positive and Gram negative bacteria and a number of fungi. Moderate susceptibility of all tested strains was observed. Determined MIC values were 0.156–5 mg mL−1 (bacterial strains) and 0.039–0.325 mg mL−1 (fungal strains). Major component of the most active oil, palmitic acid, was also tested for activity together with stearic and myristic acids.

scholarly journals Development, optimization, standardization, and validation of a simple in-house agar gradient method to determine minimum inhibitory concentration of vancomycin for Staphylococcus aureus

2019 ◽  
Vol 11 (03) ◽  
pp. 220-228
Author(s):  
Sumit Rai ◽  
Mukta Tandon ◽  
Narendra Pal Singh ◽  
Vikas Manchanda ◽  
Iqbal Rajinder Kaur
Keyword(s):  
Staphylococcus Aureus ◽  
Gradient Method ◽  
Inhibitory Concentration ◽  
Control Strain ◽  
Broth Microdilution ◽  
Clinical Strains ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Commercial Strip ◽  
Validation Stage

Abstract BACKGROUND: The Clinical and Laboratory Standards Institute recommends reporting minimum inhibitory concentration (MIC) values of vancomycin for Staphylococcus aureus. Commercial MIC strips are expensive, and the traditional broth microdilution method is cumbersome. With this background, we attempted to develop and standardize an in-house agar gradient method to determine MIC values of vancomycin for S. aureus. OBJECTIVES: To develop and validate an in-house vancomycin MIC strip, based on simple agar gradient method for S. aureus as per bioassay development guidelines. MATERIALS AND METHODS: Filter paper gradient strips were made in house and impregnated with varying concentrations of vancomycin to create an antibiotic gradient. During standardization, MICs of ninety clinical strains of S. aureus and ATCC 29213 were tested by the broth microdilution and commercial strip followed by the in-house strip. During the validation stage, MICs of ninety different clinical strains of S. aureus and ATCC 29213 were determined by the in-house strip followed by MIC detection by broth microdilution and commercial strips. A reading of more than ± 1log2 dilution compared with broth microdilution was considered as an outlier. RESULTS: During the initial stage, there were 7/90 outliers in the clinical strains, and no outliers were seen with the ATCC 29213 control strain. Corrective action included increasing precaution during the antibiotic impregnation on the strip. During validation stage, only 4/90 outliers were observed in the clinical strains. The commercial strips had 29/90 among clinical and 15/30 outliers in the control strain during the prevalidation phase. Despite maintaining cold chain during the validation phase, the outliers for commercial strip were 18/90 and 4/30 for clinical and control strains, respectively. CONCLUSION: Reporting vancomycin MIC for S. aureus may be attempted using the in-house method after validating it with a gold standard broth microdilution method and quality control as per protocol.

scholarly journals Influence of Gallic Acid and Thai Culinary Essential Oils on Antibacterial Activity of Nisin against Streptococcus mutans

2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Pimsumon Jiamboonsri ◽  
Pimpikar Kanchanadumkerng
Keyword(s):  
Essential Oils ◽  
Gallic Acid ◽  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Inhibitory Concentration ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Life Threatening ◽  
Oral Pathogens ◽  
Time Kill

Streptococcus mutans is a well-known oral pathogen commonly associated with a normal dental problem and life-threatening infection. A bacteriocin nisin and the plant-derived compounds including gallic acid (GA) and Thai culinary essential oils (EOs) have been reported to have activity against oral pathogens. However, their synergistic interaction against S. mutans has not been explored. The purposes of this study were primarily to investigate anti-S. mutans properties and the antibiofilm formation of nisin, GA, and five EOs by using the broth microdilution method. Besides, the morphological change, killing rate, and antibacterial synergism were determined by scanning electron microscopy (SEM), time-kill assay, and checkerboard method, respectively. The results demonstrated that kaffir lime leaf (KLL) oil, lemongrass (LG) oil, and GA showed a potent anti-S. mutans activity and inhibited biofilm formation with the possible mechanism targeted on the cell membrane. Additionally, KLL oil revealed anti-S. mutans synergism with GA, LG oil, and chlorhexidine with the fractional inhibitory concentration (FIC) indexes ≤ 0.5. Interestingly, GA displayed a high potential to enhance anti-S. mutans activity of nisin by lowering the minimum inhibitory concentrations (MICs) to at least 8-fold in a bacteriostatic manner. These results suggest that GA and KLL oil may be potentially used as an adjunctive therapy along with nisin and chlorhexidine to control S. mutans infection.

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